Catherwood M A, Alexander H D, McManus D T, Cuthbert R J G, Morris T C M
Department of Haematology, Belfast City Hospital, Lisburn Road, Belfast BT9 7AD, Northern Ireland, UK.
Leuk Lymphoma. 2003 Apr;44(4):645-8. doi: 10.1080/1042819021000055327.
Determination of the biologic potential of lymphoid proliferations in biopsies can be difficult by standard histological or even immunohistochemical examination. Polymerase chain reaction (PCR) has been used with increasing frequency to detect clonal rearrangements of the immunoglobulin heavy chain (IgH) in formaldehyde fixed, paraffin wax embedded tissues. Sensitivity ranges between 50 and 80%, and therefore at least 20% of neoplasms remain undetected by these approaches. Few investigators have attempted to detect immunoglobulin light chain (IgL) gene rearrangements by PCR using paraffin wax embedded samples. We studied 29 cases of B-cell neoplasms, along with 21 cases with equivocal histology and 4 reactive biopsies, using degenerate oligoprimers to amplify Ig(kappa) and Ig(lambda) light chain genes, along with IgH (Fr 1, 2 and 3) gene rearrangement analysis. The combination of these methods detected clonality in 93% of cases (27/29) with histological diagnosis of B-NHL. Fr2 and Fr3 primers detected clonality in 79% (23/29) of cases. IgL chain rearrangements detected 4 cases (14%), negative for IgH rearrangements, improving sensitivity from 79 to 93%. Clonality was detected in 52% (11/21) of histologically equivocal lymphoid proliferations, including one case detected by IgL rearrangements which was negative for IgH rearrangements. Archival material from 4 cases with reactive histology produced polyclonal results. These results confirm that PCR based immunoglobulin gene rearrangement is a sensitive and specific method for demonstrating B-cell clonality in paraffin-wax embedded sections. The addition of IgL analysis to the IgH assay allows the detection of greater than 90% of B-cell lymphoproliferative disorders from routine histological specimens with poor preservation of genomic DNA.
通过标准组织学甚至免疫组织化学检查来确定活检中淋巴样增生的生物学潜能可能具有挑战性。聚合酶链反应（PCR）已越来越频繁地用于检测甲醛固定、石蜡包埋组织中免疫球蛋白重链（IgH）的克隆重排。其灵敏度在50%至80%之间，因此至少20%的肿瘤通过这些方法仍未被检测到。很少有研究者尝试使用石蜡包埋样本通过PCR检测免疫球蛋白轻链（IgL）基因重排。我们使用简并寡核苷酸引物扩增Ig（κ）和Ig（λ）轻链基因以及IgH（框架1、2和3）基因重排分析，研究了29例B细胞肿瘤，以及21例组织学不明确的病例和4例反应性活检。这些方法的组合在93%（27/29）经组织学诊断为B-NHL的病例中检测到克隆性。框架2和框架3引物在79%（23/29）的病例中检测到克隆性。IgL链重排在4例（14%）IgH重排阴性的病例中被检测到，将灵敏度从79%提高到93%。在52%（11/21）组织学不明确的淋巴样增生中检测到克隆性，包括1例通过IgL重排检测到而IgH重排阴性的病例。4例反应性组织学的存档材料产生多克隆结果。这些结果证实基于PCR的免疫球蛋白基因重排是一种在石蜡包埋切片中证明B细胞克隆性的敏感且特异的方法。在IgH检测中加入IgL分析可使从常规组织学标本中检测出超过90%的B细胞淋巴增殖性疾病，这些标本的基因组DNA保存较差。
